UniProt UniProt http://www.uniprot.org Mob2p interacts with the protein kinase Orb6p to promote coordination of cell polarity with cell cycle progression. Mob2p interacts with the protein kinase Orb6p to promote coordination of cell polarity with cell cycle progression. J. Cell. Sci. (0021-9533) 2003 Hou MC., Wiley DJ., Verde F., McCollum D. dannel.mccollum@umassmed.edu 22-Jan-2008: Contacted by JYOTI. A phage lambda cDNA library was used. orchard in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip autoradiography autoradiography Hou MC., Wiley DJ., Verde F., McCollum D. dannel.mccollum@umassmed.edu Accepted 2007-Not a month!-11 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. A phage lambda cDNA library was used. J. Cell. Sci. (0021-9533) 2003 orchard Mob2p interacts with the protein kinase Orb6p to promote coordination of cell polarity with cell cycle progression. Mob2p interacts with the protein kinase Orb6p to promote coordination of cell polarity with cell cycle progression. J. Cell. Sci. (0021-9533) 2003 Hou MC., Wiley DJ., Verde F., McCollum D. dannel.mccollum@umassmed.edu 22-Jan-2008: Contacted by JYOTI. A phage lambda cDNA library was used. orchard yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Hou MC., Wiley DJ., Verde F., McCollum D. dannel.mccollum@umassmed.edu Accepted 2007-Not a month!-11 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. A phage lambda cDNA library was used. J. Cell. Sci. (0021-9533) 2003 orchard Mob2p interacts with the protein kinase Orb6p to promote coordination of cell polarity with cell cycle progression. Mob2p interacts with the protein kinase Orb6p to promote coordination of cell polarity with cell cycle progression. J. Cell. Sci. (0021-9533) 2003 Hou MC., Wiley DJ., Verde F., McCollum D. dannel.mccollum@umassmed.edu 22-Jan-2008: Contacted by JYOTI. A phage lambda cDNA library was used. orchard yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Hou MC., Wiley DJ., Verde F., McCollum D. dannel.mccollum@umassmed.edu Accepted 2007-Not a month!-11 by ORCHARD. A phage lambda cDNA library was used. J. Cell. Sci. (0021-9533) 2003 orchard Mob2p interacts with the protein kinase Orb6p to promote coordination of cell polarity with cell cycle progression. Mob2p interacts with the protein kinase Orb6p to promote coordination of cell polarity with cell cycle progression. J. Cell. Sci. (0021-9533) 2003 Hou MC., Wiley DJ., Verde F., McCollum D. dannel.mccollum@umassmed.edu 22-Jan-2008: Contacted by JYOTI. A phage lambda cDNA library was used. orchard schpm Schizosaccharomyces pombe (fission yeast) Fission yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Hou MC., Wiley DJ., Verde F., McCollum D. dannel.mccollum@umassmed.edu Accepted 2007-Not a month!-11 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. A phage lambda cDNA library was used. J. Cell. Sci. (0021-9533) 2003 orchard aakg_schpo 5'-AMP-activated protein kinase subunit gamma cbs2 SPAC1556.08c SPAC1F12.01c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MTDVQETQKGALKEIQAFIRSRTSYDVLPTSFRLIVFDVTLFVKTSLSLLTLNNIVSAPLWDSEANKFAGLLTMADFVNVIKYYYQSSSFPEAIAEIDKFRLLGLREVERKIGAIPPETIYVHPMHSLMDACLAMSKSRARRIPLIDVDGETGSEMIVSVLTQYRILKFISMNCKETAMLRVPLNQMTIGTWSNLATASMETKVYDVIKMLAEKNISAVPIVNSEGTLLNVYESVDVMHLIQDGDYSNLDLSVGEALLKRPANFDGVHTCRATDRLDGIFDAIKHSRVHRLFVVDENLKLEGILSLADILNYIIYDKTTTPGVPEQTDNFESAV 8D32CC6CE53F7916 8D32CC6CE53F7916 orb6_schpo Serine/threonine-protein kinase orb6 orb6 SPAC821.12 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MDKNDYLHFERNPSLFPKSTLDKVQKTKKYIEHYYKVAVDHAVERNQRRINLEQRLATERGSEERKNRQLRASGEKESQFLRFRRTRLSLEDFSTIKVIGKGAFGEVRLVQKLDTGKIYAMKSLLKTEMFKRDQLAHVKAERDLLVESDSPWVVSLYYAFQDSLYLYLIMEFLPGGDLMTMLINYDTFSEDVTRFYMAECVLAIADVHRMGYIHRDIKPDNILIDRDGHIKLSDFGLSTGFYKQDQSASYMKPRTGNTVKRGQMVDAIWLTMSSKDKMATWKKNRRVMAYSTVGTPDYIAPEIFLQQGYGQDCDWWSLGAIMFECLIGWPPFCSENSHETYRKIINWRETLTFPNDIHLSIEARDLMDRLMTDSEHRLGRGGAIEIMQHPFFTGIDWDHIRETAAPFIPNLKSITDTHYFPVDELEQVPEQPVTQQPASVDPQTLEQTNLAFLGYTYKKFNYLTMKGAL 1BFF2905E81FBF6C snf1_schpo SNF1-like protein kinase ssp2 ssp2 SPCC74.03c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MQPQEVDLMENSTMRNGARVLPPEAISKRHIGPYIIRETLGEGSFGKVKLATHYKTQQKVALKFISRQLLKKSDMHMRVEREISYLKLLRHPHIIKLYDVITTPTDIVMVIEYAGGELFDYIVEKKRMTEDEGRRFFQQIICAIEYCHRHKIVHRDLKPENLLLDDNLNVKIADFGLSNIMTDGNFLKTSCGSPNYAAPEVINGKLYAGPEVDVWSCGIVLYVMLVGRLPFDDEFIPNLFKKVNSCVYVMPDFLSPGAQSLIRRMIVADPMQRITIQEIRRDPWFNVNLPDYLRPMEEVQGSYADSRIVSKLGEAMGFSEDYIVEALRSDENNEVKEAYNLLHENQVIQEKSHLSKSKRVDSFLSVSPPAFSEYTSELQKKSKQELIDPTLEGPRWTVSDPPTYAKQTIDSNICVLVPTAEKNKLEMRTLADAASAVDTSQSTRKKSRRNKWHFGVRCRGDAPEILLAVYRALQRAGAQFTVPKPVNGKYRSDMYTIKSRWEIPHCKREGKNTYAYIELQLYEVMPGCFMLDVKSNGYKDIYSHPERTADHGMDDLKSSFPFLDLCAMLVCKLFSA E5857E8F171E7B50 E5857E8F171E7B50 mob2_schpo Maintenance of ploidy protein mob2 mob2 SPCC970.04c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MFLLNSLSRITRGNRSKRHQNLSDASSSSGSFSKKSSTSQLVRTGSPSVEPTALYLQQPFVRTHLVKGNFSTIVSLPRFVDLDEWVALNVYELFTYLNHFYDVFATFCTVKTCPVMSAAANFDYTWLDNNRKPVHLPAPQYIEYVLAWIENRLHDQNVFPTKAGLPFPSNFLVIVKAIYKQMFRIFAHMYYAHYAEILHLSLEAHWNSFFAHFIAFGKEFQLLDKRDTAPLKDLIVVLENQGNI 62630694D7BA2608 orb6-mbo2-1 3 6 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association In fact, Mob2p was identified independently in a two-hybrid screen for Orb6p-binding partners (see Materials and Methods). orb6-mbo2-2 2 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Fig 5A Beta-galactosidase assays to test for interaction between Mob2p and Orb6p snf1-snf4-3 2 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Fig 5A orb6-mbo2-3 4 6 unspecified role unspecified role bait bait c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role prey prey c-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig 5B Using a strain (YDM1151) expressing chromosomal Mob2p-13Myc and Orb6p-3HA, we found that the two proteins could be co-immunoprecipitated (Fig. 5B) mbo2-orb6-1 1 8 unspecified role unspecified role bait bait n-terminus myc his tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 6 unspecified role unspecified role prey prey n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction Fig 5C Mob2p and Orb6p were also found to interact by a binding assay using in vitro translated proteins (Fig. 5C), suggesting that their interaction is direct.